嗜热四膜虫有性后代的分离纯化

doi:10.1385.9/j.cjz.201902014

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嗜热四膜虫有性后代的分离纯化
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                        10.1385.9/j.cjz.201902014
                    
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作者单位:① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049,① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049,① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049,① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049,① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049,① 中国科学院水生生物研究所水生生物多样性与保护重点实验室 武汉 430072;② 中国科学院大学现代农业科学学院 北京 100049
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基金项目:国家自然科学基金项目（No. 91631303）；

Isolation and Purification of Sexual Progeny of Tetrahymena Thermophila

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①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049,①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049,①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049,①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049,①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049,①Key Laboratory of Aquatic Biodiversity and Conservation,Institute of Hydrobiology,Chinese Academy of Sciences,Wuhan,430072; ②College of Advanced Agricultural Sciences,University of Chinese Academy of Sciences,Beijing,100049

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摘要:

在实验条件下诱导四膜虫种群交配进行有性生殖时，交配后的种群中除有性后代外，还会混杂一定比例的亲本细胞，而高纯度的有性后代是研究四膜虫一些基本生物学问题的重要基础。本研究利用四膜虫交配时不能摄食并且新产生的有性后代几小时后才恢复摄食能力的特性，以嗜热四膜虫（Tetrahymena thermophila）交配种群为对象，在其有性生殖阶段后期向交配体系中加入Fe3O4磁性纳米颗粒，未交配的亲本细胞由于能够摄入磁性颗粒而具有磁性；再利用有性后代和未交配细胞的磁性差异，在磁场作用下将有性后代分离纯化出来。通过优化过柱分离的时间点、分离柱的口径、分离柱中铁粉/石英粉的质量等分离条件，有性后代的分离纯度及收率均能达到98%以上。该分离方法亦可为其他种四膜虫或纤毛虫有性后代的分离纯化提供借鉴。

Abstract:

When Tetrahymena cells are induced to mate for sexual reproduction in the lab, the resulting population always contains a certain proportion of unmated parental cells besides sexual progeny. However, highly purified sexual progeny is an important basis for studying some basic biological questions of Tetrahymena. In this study, based on the characteristics that mating Tetrahymena loses the ability to ingest particles and the newly produced sexual progeny does not recover this ability within several hours, we added the Fe3O4 magnetic nanoparticles to the mating population of T. thermophila at the late stage of sexual reproduction and only the unmated parent cells would become magnetic because they still had the ability to ingest the magnetic particles, which made possible an efficient isolation of sexual progeny under the action of magnetic field. By optimizing the time point at which cell population was passed through the separation column, the diameter of the column and the quality of iron/quartz powder in the column, we could make the purity and yield of the sexual progeny both more than 98%. The established method can also be used for the separation and purification of the sexual progeny of other Tetrahymena or ciliate species.

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马雪凤,孙宗毅,张晶,陈凯,王光营,缪炜.2019.嗜热四膜虫有性后代的分离纯化.动物学杂志,54(2):279-287.

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收稿日期:2018-09-16
最后修改日期:2019-02-14
录用日期:2019-02-12
在线发布日期: 2019-03-25
出版日期: 2019-04-20

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